During this Phase I SBIR project, Electronic BioSicences (EBS) will develop a direct, long-read, RNA strand sequencing methodology that is capable of sequencing both canonical and specific modified nucleotides with high-accuracy. This will result in the development of technology wherein a single-stranded RNA (ssRNA) molecule is translocated through a nanopore reader using an applied voltage bias and the sequence is directly read from the resulting translocation current signature. This methodology will be innovative relative to other nanopore-based approaches available in that no processive enzymes will be involved, yielding a new technology that is both long-read capable and high-accuracy, an accomplishment that is needed in the field of epitranscriptomics but not presently available. The resulting technology will be capable of providing a new and detailed understanding RNA and its regulation.